Preparation and electrophoretic separation of Bodipy-Fl-labeled glycosphingolipids.

The University of Notre Dame, Department of Chemistry and Biochemistry, Notre Dame, IN 46556, USA.
Journal of Chromatography A (Impact Factor: 4.61). 03/2012; 1229:268-73. DOI: 10.1016/j.chroma.2012.01.031
Source: PubMed

ABSTRACT Several glycosphingolipids were labeled with the fluorphore Bodipy-Fl and analyzed using capillary electrophoresis with laser-induced fluorescence detection. GM1-, LacCer-, and Cer-Bodipy-Fl were prepared through acylation using the N-hydroxysuccinimide ester of Bodipy-Fl. Several other glycosphingolipids including GT1a-, GD1a-, GM2-, GM3-, GD3-, and GlcCer-Bodipy-Fl were enzymatically synthesized. Micellar electrokinetic capillary chromatography with a TRIS/CHES/SDS/α-cyclodextrin buffer produced better separation than an established borate/deoxycholate/methyl-β-cyclodextrin buffer. The nine Bodipy-Fl-labeled glycosphingolipid standards were separated in under 5 min, theoretical plate counts were between 640,000 and 740,000, and the limit of detection was approximately 3 pM or 240 ymol analyte injected onto the capillary.

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